Immuno-Northern Blotting: Detection of RNA Modifications by Using Antibodies against Modified Nucleosides
Fig 4
Immuno-northern blot analysis of mitochondrial and non-mitochondrial RNA.
RNAs were isolated from the total fraction (Total), mitochondrial fraction (Mito.), and non-mitochondrial fraction (Non-mito.) of mouse liver. (A) Each total RNA (0.6 μg) was separated in the 12% polyacrylamide gel and analyzed by SYBR staining and INB with anti-m1A antibody. The red arrowheads denote shorter tRNA bands, which were rich in the mitochondrial RNA. Arrows denote slight anti-m1A positive bands in the long exposure image. White arrowheads denote negative signals in 5.8S and 5S rRNA. (B) Each total RNA (1.2 μg) was separated on the 1% agarose gel and analyzed. The arrowheads denote positive signals in 28S and 18S rRNA. The arrow probably denotes the 28S rRNA-derived degraded transcript produced during the sample preparation.